title: Isolation, characterization and differentiation of cells expressing pluripotent/multipotent markers from adult human ovaries.
abstract: Pluripotent stem cells are still generally accepted not to exist in adult human ovaries, although increasing studies confirm the presence of pluripotent/multipotent stem cells in adult mammalian ovaries, including those of humans. The aim of this study is to isolate, characterize and differentiate in vitro stem cells that originate from the adult human ovarian cortex and that express markers of pluripotency/multipotency. After enzymatic degradation of small ovarian cortex biopsies retrieved from 18 women, ovarian cell cultures were successfully established from 17 and the formation of cell colonies was observed. The presence of cells/colonies expressing some markers of pluripotency (alkaline phosphatase, surface antigen SSEA-4, OCT4, SOX-2, NANOG, LIN28, STELLA), germinal lineage (DDX4/VASA) and multipotency (M-CAM/CD146, Thy-1/CD90, STRO-1) was confirmed by various methods. Stem cells from the cultures, including small round SSEA-4-positive cells with diameters of up to 4 μm, showed a relatively high degree of plasticity. We were able to differentiate them in vitro into various types of somatic cells of all three germ layers. However, these cells did not form teratoma when injected into immunodeficient mice. Our results thus show that ovarian tissue is a potential source of stem cells with a pluripotent/multipotent character for safe application in regenerative medicine.
authors.affiliation: Reproductive Unit, Department of Obstetrics and Gynaecology, University Medical Centre Ljubljana, Slajmerjeva 3, 1000, Ljubljana, SI, Slovenia.
authors.forename: Martin
authors.initials: M
authors.lastname: Stimpfel
authors.forename: Thomas
authors.initials: T
authors.lastname: Skutella
authors.forename: Branko
authors.initials: B
authors.lastname: Cvjeticanin
authors.forename: Marija
authors.initials: M
authors.lastname: Meznaric
authors.forename: Peter
authors.initials: P
authors.lastname: Dovc
authors.forename: Srdjan
authors.initials: S
authors.lastname: Novakovic
authors.forename: Petra
authors.initials: P
authors.lastname: Cerkovnik
authors.forename: Eda
authors.initials: E
authors.lastname: Vrtacnik-Bokal
authors.forename: Irma
authors.initials: I
authors.lastname: Virant-Klun
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D008875:Middle Aged; D039902:Multipotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D010053:Ovary / Q000166:cytology* / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D055746:Stage-Specific Embryonic Antigens / Q000032:analysis
chemical_list: D055746:Stage-Specific Embryonic Antigens; C066584:stage-specific embryonic antigen-4